Interfering with Resistance to Smoothened Antagonists by Inhibition of the PI3K Pathway in Medulloblastoma

Resistance of medulloblastoma to Smo antagonists can be delayed or prevented by specific drug combinations. An End Run Against Tumor Resistance Cancer cells are as clever as microbes. Mustering their considerable abilities to rapidly replicate and evolve, both cancer cells and bacteria quickly develop resistance to the drugs we use to fight them. Modern medicine confronts a growing population of pathogens that cannot be treated by our usual antibiotics, and oncologists must be prepared with second- and third-line therapies, because tumors that retreat from initial drug treatments often return with renewed vigor. Buonamici et al. confront this problem in their study of a new class of cancer therapeutic agents now in clinical trials—antagonists of a membrane protein called Smoothened (Smo). The Smo receptor normally regulates a developmental pathway but is abnormally activated in medulloblastoma (a malignant brain tumor) and basal cell carcinoma of the skin. Medulloblastomas in mice respond well to these Smo antagonists but soon become resistant, these authors find. If, however, an inhibitor of the phosphatidylinositol 3-kinase (PI3K) signaling pathway is added to the initial drug cocktail, resistance is delayed or even prevented. In some cancers, the Smo receptor is active even when its ligand is absent, conferring dependence of the tumor on the downstream Hedgehog signaling pathway, which ultimately regulates gene expression through the Gli transcription factors. Treatment of Smo-addicted tumors in mice with Smo antagonists ultimately leads to development of resistance, although tumor growth is inhibited for a while. The authors found that the tumors eluded the drug in several ways: The genes for the Gli transcription factors were sometimes amplified, compensating for loss of pathway stimulation. In other resistant tumors, there were point mutations in the Smo receptor itself that allowed reactivation of the pathway. In yet another group of tumors, by examining which genes were up-regulated, the authors found activation of a completely different signaling pathway—the PI3K pathway. Further experiments in medulloblastoma-bearing mice revealed that resistance could be delayed or even prevented by including a PI3K inhibitor along with the Smo antagonist in the initial treatment that tumor-bearing animals received. The PI3K inhibitor alone had no effect. By looking at resistance mechanisms to Smo antagonists before the drug is used in the clinic, the results of this study will better arm oncologists against the molecular defenses that cancers may commandeer to evade this drug. And by identifying a drug combination that delays or even combats development of resistance when used as a first-line treatment in clinical trials, these results could ultimately improve the lives of patients with medulloblastoma or other cancers that depend on Smo for their survival. The malignant brain cancer medulloblastoma is characterized by mutations in Hedgehog (Hh) signaling pathway genes, which lead to constitutive activation of the G protein (heterotrimeric guanosine triphosphate–binding protein)–coupled receptor Smoothened (Smo). The Smo antagonist NVP-LDE225 inhibits Hh signaling and induces tumor regression in animal models of medulloblastoma. However, evidence of resistance was observed during the course of treatment. Molecular analysis of resistant tumors revealed several resistance mechanisms. We noted chromosomal amplification of Gli2, a downstream effector of Hh signaling, and, more rarely, point mutations in Smo that led to reactivated Hh signaling and restored tumor growth. Analysis of pathway gene expression signatures also, unexpectedly, identified up-regulation of phosphatidylinositol 3-kinase (PI3K) signaling in resistant tumors as another potential mechanism of resistance. Probing the relevance of increased PI3K signaling, we demonstrated that addition of the PI3K inhibitor NVP-BKM120 or the dual PI3K-mTOR (mammalian target of rapamycin) inhibitor NVP-BEZ235 to the initial treatment with the Smo antagonist markedly delayed the development of resistance. Our findings may be useful in informing treatment strategies for medulloblastoma.

[1]  T. Curran,et al.  Loss of p53 but not ARF accelerates medulloblastoma in mice heterozygous for patched. , 2001, Cancer research.

[2]  J. D. Benson,et al.  Contribution of Polycomb Homologues Bmi-1 and Mel-18 to Medulloblastoma Pathogenesis , 2007, Molecular and Cellular Biology.

[3]  C. Emerson,et al.  Phosphoinositide 3-kinase and Akt are essential for Sonic Hedgehog signaling. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[4]  G. Rao,et al.  Sonic hedgehog and insulin-like growth factor signaling synergize to induce medulloblastoma formation from nestin-expressing neural progenitors in mice , 2004, Oncogene.

[5]  J. Engelman,et al.  Acquired resistance to tyrosine kinase inhibitors during cancer therapy. , 2008, Current opinion in genetics & development.

[6]  Y. Benjamini,et al.  More powerful procedures for multiple significance testing. , 1990, Statistics in medicine.

[7]  M. Scott,et al.  Insulin-like growth factor 2 is required for progression to advanced medulloblastoma in patched1 heterozygous mice. , 2008, Cancer research.

[8]  Hua Tian,et al.  A paracrine requirement for hedgehog signalling in cancer , 2008, Nature.

[9]  Jeremy Stinson,et al.  Treatment of medulloblastoma with hedgehog pathway inhibitor GDC-0449. , 2009, The New England journal of medicine.

[10]  Jussi Taipale,et al.  Hedgehog: functions and mechanisms. , 2008, Genes & development.

[11]  J. Kessler,et al.  Loss of patched and disruption of granule cell development in a pre-neoplastic stage of medulloblastoma , 2005, Development.

[12]  C. Rudin,et al.  Smoothened Mutation Confers Resistance to a Hedgehog Pathway Inhibitor in Medulloblastoma , 2009, Science.

[13]  Sean Ekins,et al.  Pathway mapping tools for analysis of high content data. , 2007, Methods in molecular biology.

[14]  M. Warmuth,et al.  Discovery of NVP-LDE225, a Potent and Selective Smoothened Antagonist. , 2010, ACS medicinal chemistry letters.

[15]  Paul A. Northcott,et al.  Multiple recurrent genetic events converge on control of histone lysine methylation in medulloblastoma , 2009, Nature Genetics.

[16]  S. Baylin,et al.  Cooperation between the Hic1 and Ptch1 tumor suppressors in medulloblastoma. , 2008, Genes & development.

[17]  Daniela Gabriel,et al.  Identification and characterization of NVP-BEZ235, a new orally available dual phosphatidylinositol 3-kinase/mammalian target of rapamycin inhibitor with potent in vivo antitumor activity , 2008, Molecular Cancer Therapeutics.

[18]  R. Toftgård,et al.  Hedgehog beyond medulloblastoma and basal cell carcinoma. , 2010, Biochimica et biophysica acta.

[19]  M. Tremblay,et al.  Recent patents for Hedgehog pathway inhibitors for the treatment of malignancy , 2009, Expert opinion on therapeutic patents.

[20]  Maria Kasper,et al.  GLI transcription factors: mediators of oncogenic Hedgehog signalling. , 2006, European journal of cancer.

[21]  Helen Baines,et al.  Suppression of the Shh pathway using a small molecule inhibitor eliminates medulloblastoma in Ptc1(+/-)p53(-/-) mice. , 2004, Cancer cell.

[22]  Alice T. Loo,et al.  PTEN-deficient cancers depend on PIK3CB , 2008, Proceedings of the National Academy of Sciences.

[23]  W. Sellers,et al.  Abstract 4498: Biological characterization of NVP-BKM120, a novel inhibitor of phosphoinosotide 3-kinase in Phase I/II clinical trials , 2010 .

[24]  R. Toftgård Hedgehog signalling in cancer , 2000, Cellular and Molecular Life Sciences CMLS.

[25]  J. Williams,et al.  1-amino-4-benzylphthalazines as orally bioavailable smoothened antagonists with antitumor activity. , 2009, Journal of medicinal chemistry.

[26]  H. Wichterle,et al.  Small-molecule modulators of Hedgehog signaling: identification and characterization of Smoothened agonists and antagonists , 2002, Journal of biology.

[27]  D. Rowitch,et al.  Hedgehog and PI-3 kinase signaling converge on Nmyc1 to promote cell cycle progression in cerebellar neuronal precursors , 2004, Development.